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Introdução à Bioinformática – IQB001 Rafael Dias Mesquita [email protected] Laboratório de Bioinformática Departamento de Bioquímica Preparo de biblioteca para sequenciamento Exemplos dos kits TruSeq e Nextera da Illumina

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Page 1: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Introdução à Bioinformática – IQB001

Rafael Dias Mesquita [email protected]

Laboratório de Bioinformática

Departamento de Bioquímica

Preparo de biblioteca para sequenciamento Exemplos dos kits TruSeq e Nextera da Illumina

Page 2: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Preliminar: PCR

http://bioinfo.iq.ufrj.br/graduacao/links/08%20PCR.swf

Page 3: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Kit TruSeq Illumina

Page 4: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with
Page 5: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with
Page 6: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with
Page 7: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

DNA double strands created based on FW and RV strands

Page 8: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Strand specificity

U U U U U U This strand is NOT used as template on PCR

DNA double strand created based only on FW strand

Page 9: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Strand specificity

U U U U U U This strand is NOT used as template on PCR

DNA double strand created based only on FW strand

https://support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/samplepreps_truseq/truseq-stranded-total-rna-workflow/truseq-stranded-total-rna-workflow-reference-1000000040499-00.pdf

Page 10: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

Kit Nextera Illumina

Page 11: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with
Page 12: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna-flex.html

Kit Nextera Illumina

Page 13: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with

https://www.youtube.com/watch?v=-kTcFZxP6kM

Resumo de vários kits de preparação da amostra

Page 14: Preparo de biblioteca para sequenciamento Exemplos dos ... · DNA Insert Rd2 SP Index Rdl SP Index Rd2 SP' E. Denature and amplify for final product Library construction begins with